Avian transgenesis has served as a suitable approach to generate bioreactors for the manufacturing of recombinant proteins. Production in chicken cells comes with significant advantages over other systems including providing the human-like glycosylation on target proteins. In this regard, the oviduct-specific ovalbumin promoter has been one of the ideal candidates to drive the expression of transgenes. Previous plasmid-based studies on the regulatory sequences of ovalbumin promoter have led researchers to exploit ovalbumin regulatory elements out of their native genomic context (ex situ) to direct transgene expression in the transgenic chicken bioreactors. Although the inherent limitations on the ex situ use of ovalbumin promoter have promoted the use of native ovalbumin promoter for the expression of a transgene, generation of transgenic chicken is relatively difficult, inefficient, and time-consuming. To overcome these obstacles, in this study we show that CRISPR-mediated deletion of some distal ovalbumin promoter sequences in a non-oviduct cell can lead to the significant expression of the ovalbumin gene, and also a knocked-in reporter, in an estrogen-independent manner. These findings overcome the limitation of cloned promoters, where the promoter regulatory sequences have to be taken out of their cis context and also their native spatial nuclear organization into a plasmid.